Nanobiotechnology Department, Faculty of Bioscience, Tarbiat Modares University, Tehran, Iran; Biology Department, College of Science, University of Kerbala, Karbala, Iraq.
Nanobiotechnology Department, Faculty of Bioscience, Tarbiat Modares University, Tehran, Iran.
Int J Biol Macromol. 2022 Jun 15;210:252-260. doi: 10.1016/j.ijbiomac.2022.05.021. Epub 2022 May 7.
Valproic acid (VPA) usage in high dose is teratogen with low bioavailability. Hence to improve its efficacy and reduce its side effect it was encapsulated by the Nano liposomes and stabilized by the chitosan at different concentrations. The cellular uptake, biocompatibility, loading and encapsulation efficiency of the six-different formulations (1:1, 2:1, and 4:1 of chitosan-phospholipids: VPA), PC12 differentiation to neuron cells assays (gene-expression level by qRT-PCR) were conducted for the efficacy assessment of the Nano carriers. The encapsulation efficiency (EE) results revealed that the encapsulation of the VPA corresponds to the phospholipids dose, where 2:1 formulations showed higher encapsulating rate (64.5% for non-coated and 80% for coated by chitosan). The time monitored released of VPA also showed that the chitosan could enhance its controlled release too. The cellular uptake exhibited similar uptake behavior for both the coated and the non-coated Nano carriers and cytoplasmic distribution. We witnessed no toxicity effects, at different concentrations, for both formulations. Moreover, the results indicated that the gene expression level of SOX2, NeuroD1, and Neurofilament 200 increased from 1 to 5 folds for different genes. The qRT-PCR data were confirmed by the immunofluorescence antibodies staining, where Neurofilament 68 and SOX2 cell markers were modulated during differentiation of PC12 cells. Finally, our findings suggest promising potential for the Lip-VPA-Chit Nano carrier in inducing the differentiation of PC12 into neuron for treating neurodegenerative disorders.
丙戊酸（VPA）在高剂量下具有致畸性和低生物利用度。因此，为了提高其疗效并减少其副作用，将其包封在纳米脂质体中，并通过不同浓度的壳聚糖进行稳定。通过六种不同配方（壳聚糖-磷脂：VPA 的 1:1、2:1 和 4:1）的细胞摄取、生物相容性、载药量和包封效率，以及 PC12 向神经元细胞分化的测定（qRT-PCR 基因表达水平）来评估纳米载体的功效。包封效率（EE）结果表明，VPA 的包封与磷脂剂量相对应，其中 2:1 配方显示出更高的包封率（非壳聚糖涂层为 64.5%，壳聚糖涂层为 80%）。VPA 的时间监测释放也表明壳聚糖可以增强其控制释放。细胞摄取显示出两种纳米载体（涂层和非涂层）的相似摄取行为和细胞质分布。在不同浓度下，两种配方均未观察到毒性作用。此外，结果表明，SOX2、NeuroD1 和神经丝 200 的基因表达水平从 1 倍增加到 5 倍不等。qRT-PCR 数据得到免疫荧光抗体染色的证实，其中神经丝 68 和 SOX2 细胞标志物在 PC12 细胞分化过程中发生了调节。最后，我们的研究结果表明，Lip-VPA-Chit 纳米载体在诱导 PC12 向神经元分化以治疗神经退行性疾病方面具有很大的潜力。
